Kim Jung Min, Takemoto Naofumi, Arai Ken-ichi, Masai Hisao
Department of Cell Biology, Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, Tokyo 113-8613, Japan.
EMBO J. 2003 Oct 1;22(19):5260-72. doi: 10.1093/emboj/cdg497.
Cdc7 kinase is essential for initiation of DNA replication. Cdc7(-/-) mouse embryonic stem (ES) cells are non-viable but their growth can be rescued by an ectopically expressed transgene (Cdc7(-/-)tg). Here we report that, despite the normal growth capability of Cdc7(-/-)tg ES cells, the mice with the identical genetic background exhibit growth retardation. Concomi tantly, Cdc7(-/-)tg embryonic fibroblasts (MEFs) display delayed S phase entry and slow S phase progression. Furthermore, spermatogenesis of Cdc7(-/-)tg mice is disrupted prior to pachytene stage of meiotic prophase I. The impairment in spermatogenesis correlates with the extremely low level of Cdc7 protein in testes, and is rescued by introducing an additional allele of transgene, which results in increase of Cdc7 expression. The increased level of Cdc7 also recovers the growth of Cdc7(-/-)tg MEFs and mice, indicating that the developmental abnormalities of Cdc7(-/-)tg mice are due to insufficiency of Cdc7 protein. Our results indicate the requirement of a critical level of a cell-cycle regulator for mouse development and provide genetic evidence that Cdc7 plays essential roles in meiotic processes in mammals.
细胞分裂周期蛋白7(Cdc7)激酶对于DNA复制的起始至关重要。Cdc7基因敲除(Cdc7(-/-))的小鼠胚胎干细胞无法存活,但其生长可通过异位表达的转基因(Cdc7(-/-)tg)得以挽救。在此我们报告,尽管Cdc7(-/-)tg胚胎干细胞具有正常的生长能力,但具有相同遗传背景的小鼠却表现出生长迟缓。与此同时,Cdc7(-/-)tg胚胎成纤维细胞(MEFs)进入S期延迟且S期进程缓慢。此外,Cdc7(-/-)tg小鼠的精子发生在减数分裂前期I的粗线期之前就受到破坏。精子发生的受损与睾丸中Cdc7蛋白的极低水平相关,通过引入额外的转基因等位基因可使其得到挽救,这导致Cdc7表达增加。Cdc7水平的升高也恢复了Cdc7(-/-)tg MEFs和小鼠的生长,表明Cdc7(-/-)tg小鼠的发育异常是由于Cdc7蛋白不足所致。我们的结果表明小鼠发育需要细胞周期调节因子达到临界水平,并提供了遗传学证据,证明Cdc7在哺乳动物减数分裂过程中起重要作用。